title: Whole protein and defined CD8(+) and CD4(+) peptides linked to penetratin targets both MHC class I and II antigen presentation pathways.
abstract: Cytoplasmic delivery and cross-presentation of proteins and peptides is necessary for processing and presentation of antigens for the generation of cytotoxic T cells. We previously described the use of the 16 amino acid peptide penetratin from the Drosophila Antennapedia homeodomain (penetratin, Antp) to transport cytotoxic T lymphocyte epitopes derived from ovalbumin (OVA) or the Mucin-1 tumor-associated antigen into cells. We have now shown that penetratin covalently conjugated to OVA protein and linked in tandem to CD4(+) and/or CD8(+) T-cell epitopes from OVA-stimulated T cells in vitro (B3Z T-cell hybridoma and OT-I and OT-II T cells). The induction of these responses was directly mediated by the penetratin peptide as linking a nonspecific 16-mer peptide to OVA or mixing did not induce CD8(+) or CD4(+) T-cell responses in vitro. Furthermore, interferon (IFN)-γ-secreting CD4(+) and CD8(+) T cells were induced which suppressed B16.OVA tumor growth in C57BL/6 mice. Tumor protection was mediated by a CD8(+) T-cell-dependent mechanism and did not require CD4(+) help to protect mice 7 days after a boost immunization. Alternatively, 40 days after a boost immunization, the presence of CD4(+) help enhanced antigen-specific IFN-γ-secreting CD8(+) T cells and tumor protection in mice challenged with B16.OVA. Long-term CD8 responses were equally enhanced by antigen-specific and universal CD4 help. In addition, immunization with AntpOVA significantly delayed growth of B16.OVA tumors in mice in a tumor therapy model.
authors.affiliation: Immunology and Vaccine Laboratory, Centre for Immunology, Burnet Institute, Melbourne, Victoria, Australia.
authors.forename: Dodie S
authors.initials: DS
authors.lastname: Pouniotis
authors.forename: Sandra
authors.initials: S
authors.lastname: Esparon
authors.forename: Vasso
authors.initials: V
authors.lastname: Apostolopoulos
authors.forename: Geoffrey A
authors.initials: GA
authors.lastname: Pietersz
mesh_terms: D000818:Animals; D051943:Antennapedia Homeodomain Protein / Q000276:immunology / Q000378:metabolism; D017951:Antigen Presentation; D000951:Antigens, Neoplasm / Q000276:immunology; D015704:CD4 Antigens / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D016827:CD8 Antigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D004330:Drosophila; D029721:Drosophila Proteins / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D007371:Interferon-gamma / Q000096:biosynthesis / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018396:Mucin-1 / Q000276:immunology / Q000378:metabolism; D009369:Neoplasms / Q000276:immunology*; D010047:Ovalbumin / Q000276:immunology / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology
chemical_list: D051943:Antennapedia Homeodomain Protein; D000951:Antigens, Neoplasm; D015704:CD4 Antigens; D016827:CD8 Antigens; D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D029721:Drosophila Proteins; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D018396:Mucin-1; D007371:Interferon-gamma; D010047:Ovalbumin; C414409:penetratin